Difference between revisions of "Part:BBa K2442403"
 
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<partinfo>BBa_K2442403 short</partinfo>
 
<partinfo>BBa_K2442403 short</partinfo>
  
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====This subproject is discussed in full detail on our [http://2017.igem.org/Team:Glasgow/ANDGate AND-Gate] wiki page.====
  
 
[[Image:Glasgow_gfp_gate.png|450px|thumb|center|A schematic representation of a final split-GFP AND-gate]]
 
[[Image:Glasgow_gfp_gate.png|450px|thumb|center|A schematic representation of a final split-GFP AND-gate]]
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This is a composite of the GFP N-term split subunit (GFP1) [https://parts.igem.org/Part:BBa_K1789003 K1789003] and the well characterised double-terminator construct [https://parts.igem.org/Part:BBa_B0015 B0015].
 
This is a composite of the GFP N-term split subunit (GFP1) [https://parts.igem.org/Part:BBa_K1789003 K1789003] and the well characterised double-terminator construct [https://parts.igem.org/Part:BBa_B0015 B0015].
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This N-term GFP part was found to contain an extra A nucleotide immediately preceding the start-codon ATG. This erroneous nucleotide is present in the parts-registry sequencing for K1789003, but is not reported as an error. Unfortunately, now both split-GFP modules might be affected by improperly spaced ribosome binding sites. Despite this we proceeded to the next assembly stage.
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[[Image:Glasgow_ANDgate_Figure_4.png|450px|thumb|center|A Biobrick prefix error in GFP N-term part.</b> Alignment of a sequencing read of the split GFP N-terminal to a reference sequence plasmid map. The base highlighted in red indicate an insertion has occurred at those positions relative to the reference sequence. The single-A insertion may correspond to the use a taq-polymerase during the original amplification of this coding sequence. Alignments performed in ApE.]]
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Latest revision as of 03:28, 2 November 2017

GFP split N-term (GFP2) + B0015 double terminator

This subproject is discussed in full detail on our [http://2017.igem.org/Team:Glasgow/ANDGate AND-Gate] wiki page.

A schematic representation of a final split-GFP AND-gate


This is a composite of the GFP N-term split subunit (GFP1) K1789003 and the well characterised double-terminator construct B0015.


This N-term GFP part was found to contain an extra A nucleotide immediately preceding the start-codon ATG. This erroneous nucleotide is present in the parts-registry sequencing for K1789003, but is not reported as an error. Unfortunately, now both split-GFP modules might be affected by improperly spaced ribosome binding sites. Despite this we proceeded to the next assembly stage.

A Biobrick prefix error in GFP N-term part.</b> Alignment of a sequencing read of the split GFP N-terminal to a reference sequence plasmid map. The base highlighted in red indicate an insertion has occurred at those positions relative to the reference sequence. The single-A insertion may correspond to the use a taq-polymerase during the original amplification of this coding sequence. Alignments performed in ApE.


Usage and Biology

this composite was used for assembly of larger composite split-GFP AND-gate constructs.

  • K2442405 [split GFP module 1] tetR repressible promoter + GFP N-term + RBS + double terminator
  • K2442408 Full split-GFP AND-gate circuit [Split GFP module 1 + Split GFP module 2]


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]